A network pharmacology approach to determine the underlying mechanisms of action of Yishen Tongluo formula for the treatment of oligoasthenozoospermia.

Oligoasthenozoospermia is a complex disease caused by a variety of factors, and its incidence is increasing yearly worldwide. Yishen Tongluo formula (YSTLF), created by Professor Sun Zixue, has been used to treat oligoasthenozoospermia in clinical practice for several decades with a good therapeutic effect. However, the chemical and pharmacological profiles of YSTLF remain unclear and need to be elucidated. In this study, a network pharmacology approach was applied to explore the potential mechanisms of YSTLF in oligoasthenozoospermia treatment. All of the compounds in YSTLF were retrieved from the corresponding databases, and the bioactive ingredients were screened according to their oral bioavailability (OB) and drug-likeness (DL). The potential proteins of YSTLF were obtained from the traditional Chinese medicine systems pharmacology (TCMSP) database and the Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM) database, while the potential genes of oligoasthenozoospermia were obtained from the GeneCards database and the DisGeNET database. The STRING database was used to construct an interaction network according to the common targets identified by the online tool Venny for YSTLF and oligoasthenozoospermia. The topological characteristics of nodes were visualized and analyzed through Cytoscape. Biological functions and significant pathways were determined and analyzed using the Gene Ontology (GO) knowledgebase, the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Metascape. Finally, the disease-formula-compound-target-pathway network was constructed by Cytoscape. A total of 106 bioactive ingredients and 134 potential targets from YSTLF were associated with oligoasthenozoospermia or considered to be therapeutically relevant. Pathway analysis indicated that the PI3K/Akt, MAPK and apoptosis signaling pathways were significant pathways involved in oligoasthenozoospermia. In conclusion, the current study expounded the pharmacological actions and molecular mechanisms of YSTLF in treating oligoasthenozoospermia from a holistic viewpoint. The potential molecular mechanisms were closely related to antioxidative stress, antiapoptosis and anti-inflammation, with TNF, CCND1, ESR1, NFKBIA, NR3C1, MAPK8, and IL6 being possible targets. This network pharmacology prediction may offer a helpful tool to illustrate the molecular mechanisms of the Chinese herbal compound YSTLF in oligoasthenozoospermia treatment.

Effect of Trolox on sperm quality in normozospermia and oligozospermia during cryopreservation.

This study evaluated the effects of different concentrations of Trolox supplementation to cryoprotective agent (CPA) on post-thaw apoptosis-like events that include translocation of phosphatidyl serine (PS) to the cell surface, alterations in mitochondrial membrane potential (MMP), and DNA integrity of normozoospermic and oligoozoospermic semen samples. Spermatozoa from 20 normozoospermic men and 20 patients with oligoozoospermia were cryopreserved with cryo-protective agent containing 0, 20, 40, and 80 µM Trolox. Pre-cryopreservation and post-thaw sperm MMP, PS externalization and DNA fragmentation were evaluated by flow cytometry. Sperm frozen in extender with Trolox had greater MMP, lower DNA fragmentation and externalization of PS in both groups, though the most effective dose of Trolox in normozoospermic and oligoozoospermic semen samples were different. These findings support the use of Trolox as freezing extender supplement to improve the quality of cryopreserved human sperm, measured in terms of early apoptosis changes and DNA integrity, in both normozoospermic and oligoozoospermic men.

Lifestyle factors and sperm aneuploidy.

Different environmental and lifestyle factors may interfere with the normal disjunction of sister chromatids/chromosomes during meiosis and may cause aneuploidy. The aim of the study was to examine the association between lifestyle factors and sperm aneuploidy. The study population consisted of 212 healthy men under 45 years of age attending an infertility clinic for diagnostic purposes and who had a normal semen concentration of 20-300×106mL or slight oligozoospermia (semen concentration of 15-20×106/mL). All participants were interviewed and provided a semen sample. Sperm aneuploidy was assessed using multicolor FISH (DNA probes specific for chromosomes X, Y, 18, 13, 21). Results from the study suggest that lifestyle factors are related to sperm aneuploidy. A positive relationship was found between coffee drinking everyday and the lack of chromosome X or Y, as well as coffee drinking 1-6 times per week and additional chromosome 18. Wearing boxer shorts decrease the copy number changes in the whole chromosome 18, the number of additional chromosome 18 and the lack of chromosome 13. Additionally, obesity (BMI 30-40 kg/m²) was positively associated with additional chromosome 21 after being adjusted for potential confounders. These findings demonstrate that changing the men's lifestyle habits may contribute to reduction of the incidence of sperm aneuploidy. It is necessary that men continue to follow sensible health advice concerning excess weight, coffee drinking and wearing tight fitting underwear. As this is the first such study to examine different lifestyle factors and sperm aneuploidy, the results need to be confirmed on larger population.

The micronutrient supplements, zinc sulphate and folic acid, did not ameliorate sperm functional parameters in oligoasthenoteratozoospermic men.

We investigated the effects of folic acid and zinc sulphate supplementation on the improvement of sperm function in subfertile oligoasthenoteratozoospermic (OAT) men. Eighty-three OAT men participated in a 16-week intervention randomised, double-blind clinical trial with daily treatment of folic acid (5 mg day(-1) ) and zinc sulphate (220 mg day(-1) ), or placebo. Before and after treatment, semen and blood samples were obtained for determining sperm concentration, motility, and morphology, sperm viability, sperm mitochondrial function, sperm chromatin status using toluidine blue, aniline blue, acridine orange and chromomycin A3 staining; and semen and blood folate, zinc, B12 , total antioxidant capacity (TAC) and malondialdehyde (MDA) concentrations. Sperm concentration (×10(6)  ml(-1) ) increased in subfertile men receiving the combined treatment of folic acid and zinc sulphate and also in the group receiving only folic acid treatment; however, it was not statistically significant (P = 0.056 and P = 0.05, respectively). Sperm chromatin integrity (%) increased significantly in subfertile men receiving only zinc sulphate treatment (P = 0.048). However, this improvement in sperm quality was not significant after adjusting placebo effect. This study showed that zinc sulphate and folic acid supplementation did not ameliorate sperm quality in infertile men with severely compromised sperm parameters, OAT. Male infertility is a multifactorial disorder, and also nutritional factors play an important role in results of administration of supplementation on sperm parameters. However, these results should be confirmed by multiple studies in larger populations of OAT men.

[Wuzi yanzong pills increases sperm mitochondrial membrane potential and protects its ultrastructure in oligo-asthenozoospermia model rats].

OBJECTIVE: To study the effects of Wuzi Yanzong Pills (WYP) on sperm mitochondrial membrane potential (MMP) and its ultrastructure in oligo-asthenozoospermia model rats. METHODS: Oligo-asthenozoospermia models were made in 50 male rats weighing 200 - 220 g by intragastric administration of Tripterygium Glucosides at 30 mg per kg per d for 8 weeks, and then equally allocated to a model control, a Huangjing Zanyu Capsule (HZC) control, a low-dose WYP, a medium-dose WYP, and a high-dose WYP group. Another 10 age-matched normal male rats were included as normal controls. The rats in the model and normal control groups were given intragastrically distilled water at 10 ml/kg, those in the HZC group administered HZC at 3.01 g/kg, and those in the low-, medium- and high-dose WYP groups medicated with WYP at 2.30, 4.60 and 9.20 g/kg, respectively, once daily for 30 days. At 30 minutes after the last administration, we detected the sperm MMP by JC-1 fluorescent staining and flow cytometry, and examined the sperm ultrastructure under the JEM-1230 transmission electron microscope. RESULTS: JC-1 + % and its fluorescence intensity were (33.77 +/- 6.19)% and 1 468 +/- 496 in the model control, (56.34 +/- 10.35)% and 3 277 +/- 895 in the HZC control, (40.80 +/- 10.40)% and 2 016 +/- 767 in the low-dose WYP, (59.40 +/- 6.51)% and 3 897 +/- 643 in the medium-dose WYP, and (60.71 +/- 7.81)% and 3 371 +/- 647 in the high-dose WYP group, significantly reduced in comparison with (70.80 +/- 4.92)% and 4 360 +/- 945 in the normal control group (P < 0.05), but remarkably higher in the medium- and high-dose WYP groups than in the model controls (P < 0. 05). After modeling, the sperm membrane was loose and degenerated, the mitochondria swelling, variously sized and with incomplete membrane, and the axonemal structure unclear or ruptured. After 30 days of WYP administration, compared with the model control group, the rats exhibited integrated sperm membrane and mitochondrial membrane, reduced mitochondrial swelling and basically normal axonemal and microtubular structures. CONCLUSION: Tripterygium Glucosides could decrease the sperm mitochondrial membrane potential and damage the mitochondrial structure, while WYP could significantly increase the sperm mitochondrial membrane potential and reduce the sperm mitochondrial structure damage. The protection of the integrity of sperm mitochondrial structure and function is one of the mechanisms of WYP acting on oligo-asthenozoospermia.

[Effects of yijing recipe on sperm apoptosis and mitochondrial membrane potential in patients with idiopathic oligoathenoteratospermia].

OBJECTIVE: To investigate the effects of Yijing Recipe on sperm apoptosis and mitochondrial membrane potential (MMP) in patients with idiopathic oligoathenoteratospermia. METHODS: Using the self-control method, we examined sperm apoptosis and MMP in 30 patients with oligoathenoteratospermia before and after treated with Yijing Recipe. RESULTS: The rates of early sperm apoptosis (AV +/PI -) and MMP loss were significantly reduced after treatment as compared with pre-medication ([2.86 +/- 1.47]% vs [4.26 +/- 2.79]% and [21.77 +/- 13.46]% vs [41.73 +/- 20.30]%, P<0.05). No statistically significant difference was observed in the sperm death rate (PI+) before and after treatment ([34.10 +/- 16.26]% vs [30.21 +/- 13.50]%, P>0.05). CONCLUSION: Yijing Recipe can reduce early sperm apoptosis and improve MMP, which may be one of the mechanisms underlying its efficacy on oligoathenoteratospermia.

The effects of hyperbaric oxygen treatment on vigility of spermatozoids: preliminary report.

The aim of this study was to evaluate the effect of hyperbaric oxygen treatment (HBO) to the vigilance of spermatozoids. The sperm samples were obtained from the patients (n=10) with diagnosed oligospermia. The motility of spermatozoids in tested samples was not lower than 30% but it did not exceed the critical proportion of 50%. Three groups of samples were formed, with respectively 10 samples in each group. All of them were exposed to HBO conditions of 2.5 ATA for 90 minutes. Group I was exposed to HBO conditions without any addition of medium. Group II was added 5 ml of EBSS solution. Tyrode's solution in volume of 5 ml was added to group III. The evaluation of functional characteristics of sperm was carried out 30 minutes after completion of HBO treatment. Statistically significant increased in motor activity was registered in three all groups (p<0.05). The biggest increased in vigilance of spermatozoids was found in the samples of the group III (p<0.01), exceeding the critical proportion of 50% in all of the samples. The obtained results suggest that the acute exposure of the sperm samples to HBO has favorable impact to functional capacity of spermatozoids in view of their better motility.

The therapeutic effect of tissue cultured root of wild Panax ginseng C.A. Mayer on spermatogenetic disorder.

This study examined the possibility of using a tissue cultured root of wild Panax ginseng (tcwPG) as a fertility agent. The effect of tcwPG on spermatogenesis was studied using male rats. The tcwPG crude powder was administered orally to 7-week-old rats over a 6-week period. The number of sperm in the testes and epididymides was significantly higher than the control. A histological examination did not reveal any morphological changes in the testes from the tcwPG powder treated rats. Moreover, there were no significant differences in the weights of the heart, spleen, liver, kidney, brain, testes and epididymides. Oligospermia was also induced by administering 2,3,7,8-tetrachlorodaibenzo-p-dioxin (TCDD) to the rats in order to estimate the feasibility of using tcwPG as treatment for infertility caused by spermatogenic disorders. After exposing the rats to TCDD, the tcwPG saponin fraction treated rats showed some improvement in the body weight, sperm number and testis morphology. It was estimated that tcwPG had feasibility as a therapeutic agent on spermatogenic disorder.

[Experimental study on the effects of jujingwan on oligospermia].

OBJECTIVE: To investigate the effects of Jujingwan on the spermatozoal ultrastructure and apoptosis of germ cells in oligospermia patients. METHODS: We treated 50 oligospermia patients with Jujingwan and observed the spermatozoal ultrastructure, the apoptosis of germ cells and the changes in the DNA ploidy proportion of spermatogenic cells by electron microscopy and FCM before the treatment and 3, 6, 9 and 12 months after it. RESULTS: Jujingwan increased sperm acrosome base density 6 months after the treatment and remarkably improved the integrity of acrosome membrane 12 months after it, with no obvious pathological changes in the nuclei and tails. Three months after the treatment, cell debris and apoptotic cells decreased significantly as compared with pre-treatment (P < 0. 05) , and very significantly 12 months after the treatment (P <0. 01). The proportion of haploid spermatozoa increased very significantly (P <0.01) , and the lost primary spermatocytes decreased significantly (P <0. 05) compared with pre-treatment. CONCLUSION: Jujingwan can increase the density of sperm acrosome base and improve the pathological changes of acrosome membrane in oligospermia patients; it can improve the activity of acrosome enzyme and the integrity of acrosome membrane, decrease the apoptosis rate of germ cells and sperm and increase the percentage of haploid spermatozoa; it can also reduce the percentage of apoptotic bodies and diploid sperm cells. It is indicated that Jujingwan can inhibit the apoptosis of germ cells and sperm and improve spermatogenesis in oligospermia patients.

Di-(2-ethylhexyl) phthalate induces severe aspermatogenesis in mice, however, subsequent antioxidant vitamins supplementation accelerates regeneration of the seminiferous epithelium.

Di-(2-ethylhexyl) phthalate (DEHP), now regarded as an endocrine disruptor, can experimentally induce spermatogenic disturbance in laboratory animals. Our previous study demonstrated that antioxidant vitamins (vitamins C and E) supplementation during DEHP-treatment significantly protected the rat seminiferous epithelium from DEHP-gonadotoxicity. In the present study, we gave these antioxidant vitamins to mice already having fully developed aspermatogenesis because of DEHP to determine whether or not the vitamins can cure the injured seminiferous epithelium. CD-1 male mice were fed on a DEHP-containing diet for 15 days and then fed on the DEHP-free normal diet with or without supplementation of vitamins C and E in drinking water for another 50 days. The results showed that severe aspermatogenesis was induced by the DEHP-treatment but that the damaged seminiferous epithelium spontaneously recovered whether the vitamins were provided or not. This indicates that the DEHP-induced aspermatogenesis was reversible. However, the supplementation of antioxidant vitamins significantly accelerated regeneration of the injured seminiferous epithelium, suggesting that the vitamins have a therapeutic effect on DEHP-induced aspermatogenesis.

[Bilateral microsurgical varicocelectomy in infertile men]. / Mikrokhirurgicheskaia dvukhstoronniaia varikotseléktomiia u muzhchin, stradaiushchikh besplodiem.

UNLABELLED: We assessed how bilateral microsurgical varicocelectomy alters semen quality in a large cohort of infertile men with bilateral varicocele. By means of bilateral testicular biopsy, we have investigated the spermatogenesis failure in several biological men groups and determined whether it can change the patient's candidacy for assisted reproductive technology procedures (ART). MATERIALS AND METHODS: From 1993 until 2003 in the urology and andrology department of St. Antoine hospital (1993-1994), the Tenon hospital (1994-2003) and hospital Saint Louis (1997-2003), 956 patients with varicocele were consulted and operated for primary or secondary sterility. Bilateral varicocele was observed in 219 patients. Our study included 198 patients with clinical palpable and infraclinical (ultrasonic doppler scanning) bilateral varicocele who underwent microsurgical bilateral varicicolectomy with bilateral testicular biopsy. Mean patient age was 34.5 (range 19 to 65) years. Varicocele has been classified into 4 stages. All the patients underwent a complete physical examination, including supine and standing scrotal examination, as well as testicular volume determination using a standard orchidometer. Complementary investigation included: the hormone status (FSH, LH, testosterone), semen analyses (carnitine, LDHx, fructoze, zinc). On the basis of the total motile sperm count 198 patients were divided into 5 biological groups: 10 azoospermia and oligospermia extreme (0-1) x 10(6)/cc--83 patients (41.9%); 2) oligospermia severe (1.1-5) x 10(6)/cc)--37 patients (18.7%); 3) oligospermia moderate (5.1-10) x 10(6)/cc)--36 patients (18.9%). To measure the sperm quality, the spermogram was studied 3, 6 and 12 months after the operation. On the basis of testicular biopsy we defined the following spermatogenesis groups: normal spermatogenesis--9 cases, hypospermatogenesis--141 cases, maturation arrest (early-6, late-20), Sertli cell only syndrome (SCOS)--17, tubular and peritubular sclerosis--5. The correlative analysis between the FSH classes and azoo- and oligospermic subsets (83 patients) shows that azoo-oligospermic area as a whole represents only 2 patients (2.4%) in the low FSH class, 46 (55.4%) in the normal FSH range class II, 39 (46.9%) patients in the high FSH class III. All the patients were divided into 4 groups according to the type of ART for which they qualified including 0 to 1.5 million/ml (intracytoplasmatic sperm injection-ICSI)--79 patients, 1.5 to 5 million/ml (in vitro fertilization-IVF)--33 patients, 5 to 20 million/ml (intrauterine insemination--IUI)--47 patients and 20 million/ml or greater sperm (spontaneous pregnancy candidates)--39 patients. RESULTS: Postoperative sperm concentration increased significantly compared to the one before the varicocelectomy. In the group of our patients the results were the following: of 198 patients 105 (53.1%) improved the spermogram, 44 (22.2%) worsened the spermogram, the spermogram was not changed in 49 (24.8%). In the first biological group of 83 patients, 42 (50.6%) improved the spermogram that enables using several methods for this difficult group (ICSI, IVF) for assisted reproductive technique (ART). The total amount of motile sperm per ejaculation increased from (0.09 +/- 0.030) x 10(6) before varicocelectomy up to (8.9 +/- 1.3) x 10(6) afterwards. In this group the testicular biopsy shows different variants of spermatogenesis disturbances, respectively. As for biological groups 3, 4 and 5, the observed spermogram improvements sharply increased the patients' liability to spontaneous pregnancy and make them intrauterine insemination candidates. CONCLUSIONS: Microscopic varicocelectomy has minimal morbidity and recurrence and may be beneficial for certain patients. Making simultaneous biopsy in patients with azoospermia and severe oligospermia makes it possible to distinctly define the extent of spermatogenesis disturbances and to elaborate adequate treatment policy. The drastic improvement of the sperm quality after varicocelectomy increases the ART chances for spontaneous pregnancy.

Leydig cell-derived heme oxygenase-1 regulates apoptosis of premeiotic germ cells in response to stress.

Stress-induced downregulation of spermatogenesis remains poorly understood. This study examined the induction of heme oxygenase-1 (HO-1), a carbon monoxide-generating inducible enzyme, in modulation of spermatogenesis. Rats were exposed to cadmium chloride (CdCl(2)), a stressor causing oligozoospermia, and HO-1-induction was monitored by following HO isozyme expression. CdCl(2)-treated testes increased HO-1 activity and suppressed microsomal cytochromes P450, which are required for steroidogenesis. CdCl(2)-elicited HO-1 occurred mostly in Leydig cells and coincided with CO generation, as judged by bilirubin-IXalpha immunoreactivity. Under these circumstances, germ cells in peripheral regions of seminiferous tubules exhibited apoptosis; laser flow cytometry revealed that these apoptotic cells involve diploid and tetraploid germ cells, suggesting involvement of spermatogonia and primary spermatocytes in CdCl(2)-elicited apoptosis. Pretreatment with zinc protoporphyrin-IX, an HO inhibitor, but not copper protoporphyrin-IX, which does not block the enzyme, attenuated the CdCl(2)-induced apoptosis. Such antiapoptotic effects of zinc protoporphyrin-IX were repressed by supplementation of dichloromethane, a CO donor. Upon CdCl(2)-treatment, both Sertoli cells and the germ cells upregulated Fas ligand; this event was also suppressed by zinc protoporphyrin-IX and restored by dichloromethane. Thus, Leydig cells appear to use HO-1-derived CO to trigger apoptosis of premeiotic germ cells and thereby modulate spermatogenesis under conditions of stress.

In-vitro differentiation of germ cells from frozen testicular biopsy specimens.

In some men with germ cell maturation arrest, spermatogenesis can be resumed during in-vitro culture of testicular biopsy samples. In this study, we examined whether similar differentiation events can be induced in cultured germ cells from cryopreserved testicular biopsy specimens. Fresh and cryopreserved aliquots of the same testicular biopsy samples were cultured in medium supplemented with FSH and testosterone. After 24 and 48 h of culture, the progression of spermatogenesis and the percentage of Sertoli cells with DNA damage, detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL), were evaluated. Spermatogenesis progressed in a similar way in fresh and cryopreserved aliquots over the first 24 h of culture. However, in contrast to fresh aliquots, no additional progress of spermatogenesis was detected between the 24 and 48 h time points. The percentage of TUNEL-positive Sertoli cells in fresh aliquots showed only a moderate increase after 24 h of culture, whereas most Sertoli cells from cryopreserved aliquots became TUNEL-positive during the same culture period. These data show that limited progression of spermatogenesis can be achieved by culturing cryopreserved testicular biopsy specimens for 24 h, but no additional benefit can be expected from prolonging the culture beyond this time point.

Does acupuncture treatment affect sperm density in males with very low sperm count? A pilot study.

Classic therapies are usually ineffective in the treatment of patients with very poor sperm density. The aim of this study was to determine the effect of acupuncture on these males. Semen samples of 20 patients with a history of azoospermia were examined by light microscope (LM) and scanning electron microscope (SEM), with which a microsearch for spermatozoa was carried out. These examinations were performed before and 1 month after acupuncture treatment and revealed that the study group originally contained three severely oligoteratoasthenozoospermic (OTA), two pseudoazoospermic and 15 azoospermic patients. The control group was comprised of 20 untreated males who underwent two semen examinations within a period of 2-4 months and had initial andrological profiles similar to those of the experimental group. No changes in any of the parameters examined were observed in the control group. There was a marked but not significant improvement in the sperm counts of severely OTA males following acupuncture treatment (average = 0.7 +/- 1.1 x 10(6) spermatozoa per ejaculate before treatment vs. 4.3 +/- 3.2 x 10(6) spermatozoa per ejaculate after treatment). A definite increase in sperm count was detected in the ejaculates of 10 (67%) of the 15 azoospermic patients. Seven of these males exhibited post-treatment spermatozoa that were detected even by LM. The sperm production of these seven males increased significantly, from 0 to an average of 1.5 +/- 2.4 x 10(6) spermatozoa per ejaculate (Z = -2.8, P < or = 0.01). Males with genital tract inflammation exhibited the most remarkable improvement in sperm density (on average from 0.3 +/- 0.6 x 10(6) spermatozoa per ejaculate to 3.3 +/- 3.2 x 10(6) spermatozoa per ejaculate; Z = -2.4, P < or = 0.02). Two pregnancies were achieved by the IVF-ICSI procedure. It is concluded that acupuncture may be a useful, nontraumatic treatment for males with very poor sperm density, especially those with a history of genital tract inflammation.

Antioxidant treatment of patients with asthenozoospermia or moderate oligoasthenozoospermia with high-dose vitamin C and vitamin E: a randomized, placebo-controlled, double-blind study.

In a randomized, placebo-controlled, double-blind study we investigated whether high-dose oral treatment with vitamins C and E for 56 days was able to improve semen parameters of infertile men. Ejaculate parameters included semen volume, sperm concentration and motility, and sperm count and viability. Thirty-one patients without genital infection but with asthenozoospermia (< 50% motile spermatozoa) and normal or only moderately reduced sperm concentration (> 7 x 10(6) spermatozoa/ml) (according to WHO criteria) were examined. To investigate the influence of the epididymal storage period on semen parameters, the patients were asked to deliver two semen samples with abstinence times of 2 and 7 days both before and at the end of vitamin treatment. After randomization, the patients received either 1000 mg vitamin C and 800 mg vitamin E (n = 15) or identical placebo capsules (n = 16). No changes in semen parameters were observed during treatment, and no pregnancies were initiated during the treatment period. Combined high-dose antioxidative treatment with vitamins C and E did not improve conventional semen parameters or the 24-h sperm survival rate. Prolonged abstinence time increased ejaculate volume (P < 0.05), sperm count (P < 0.05), sperm concentration (P < 0.05) and the total number of motile spermatozoa (P < 0.05).

An improved treatment procedure for testicular biopsy specimens offers more efficient sperm recovery: case series.

OBJECTIVE: To report an improved sperm recovery procedure from testicular biopsy specimens for intracytoplasmic sperm injection (ICSI). DESIGN: Case series and controlled study. SETTING: Procedures were performed in a tertiary IVF center. PATIENT(S): Nonobstructive azoospermic cases (15 patients) and obstructive azoospermic cases (5 patients). INTERVENTION(S): Intracytoplasmic sperm injection was carried out using testicular sperm isolated from a testicular biopsy specimen either with or without erythrocyte lysing buffer treatment. MAIN OUTCOME MEASURE(S): The time required to collect spermatozoa and the intactness and fertilization and developmental rates of oocytes. RESULT(S): In 7 of the 15 nonobstructive cases, it was possible to perform ICSI when, after shredding of the testicular tissue, no (or virtually no) sperm were present. There was no difference in the fertilization rates (83% and 68%) and developmental rates (87% and 89%) of the 54 sibling oocytes from another 5 patients in whom ICSI was carried out with sperm either treated or not treated with erythrocyte lysing buffer. CONCLUSION(S): Erythrocyte lysing buffer treatment of testicular biopsy specimens enhances the efficiency of sperm collection in those cycles in which spermatozoa are present and does not affect fertilization and embryo development.

Live birth resulting from in-office vas deferens sperm retrieval combined with natural-cycle insemination.

In-office retrieval of spermatozoa from the vas deferens was successfully combined with intra-uterine insemination of the wife during her natural cycle in the case of a man with azoospermia due to a non-correctable obstruction of the vas deferens. The combined cost of both procedures (sperm retrieval and intra-uterine insemination with processed spermatozoa) was less than $1000. The cost and convenience compare quite favourably with that of epididymis sperm aspiration combined with in-vitro fertilization, which is the currently recommended alternative treatment in such cases. These alternative therapies are compared and discussed relative to their advantages and disadvantages. Our technique for in-office vas deferens sperm retrieval is also presented.